| Abstract: | Abstract A simple, accurate method combining low pressure liquid affinity chromatography and flow injection analysis is described for the quantitation of immunoglobulins in biologicals fluids. The affinity matrix consists of Protein A covalently immobilized to a 2-fluoro-1 methylpyridinium salt activated Fractogel support. Utilizing a 1 cm3 affinity column, optimized binding and eluting buffer flow rates of 0.7 and 1.5 mL min?1, respectively, and a sample loop size of 100 μL, IgG's can be eluted from the affinity column in about 9 min. Linear standard curves (r > 0.99) were obtained at concentrations up to at least 4.0 and 8.0 mg mL?1 respectively for human and bovine IgG. Recovery yields are good ranging from 96–100%. The within day CV for human and bovine IgG was found to be less than 3.0% whereas the day to day CV was less than 3.4% (n=10). IgG concentrations of spiked and unspiked bovine plasma samples obtained by the low pressure affinity/flow injection method when compared to those obtained by the radial immunodiffusion agreed to within 4%. |
